Calpain regulates enterocyte brush border actin assembly and pathogenic Escherichia coli-mediated effacement

David A. Potter, Anjaiah Srirangam, Kerry A. Fiacco, Daniel Brocks, John Hawes, Carter Herndon, Masatoshi Maki, David Acheson, Ira M. Herman

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

This study identifies calpain as being instrumental for brush border (BB) microvillus assembly during differentiation and effacement during bacterial pathogenesis. Calpain activity is decreased by 25-80% in Caco 2 lines stably overexpressing calpastatin, the physiological inhibitor of calpain, and the effect is proportional to the calpastatin/calpain ratio. These lines exhibit a 2.5-fold reduction in the rate of microvillus extension. Apical microvillus assembly is reduced by up to 50%, as measured by quantitative fluorometric microscopy (QFM) of ezrin, indicating that calpain recruits ezrin to BB microvilli. Calpain inhibitors ZLLYCHN2, MDL 28170, and PD 150606 block BB assembly and ezrin recruitment to the BB. The HIV protease inhibitor ritonavir, which inhibits calpain at clinically relevant concentrations, also blocks BB assembly, whereas cathepsin and proteasome inhibitors do not. Microvillus effacement is inhibited after exposure of calpastatin-overexpressing cells to enteropathogenic Escherichia coli. These results suggest that calpain regulates BB assembly as well as pathological effacement, and indicate that it is an important regulator involved in HIV protease inhibitor toxicity and host-microbial pathogen interactions.

Original languageEnglish (US)
Pages (from-to)30403-30412
Number of pages10
JournalJournal of Biological Chemistry
Volume278
Issue number32
DOIs
StatePublished - Aug 8 2003

Fingerprint Dive into the research topics of 'Calpain regulates enterocyte brush border actin assembly and pathogenic Escherichia coli-mediated effacement'. Together they form a unique fingerprint.

  • Cite this

    Potter, D. A., Srirangam, A., Fiacco, K. A., Brocks, D., Hawes, J., Herndon, C., Maki, M., Acheson, D., & Herman, I. M. (2003). Calpain regulates enterocyte brush border actin assembly and pathogenic Escherichia coli-mediated effacement. Journal of Biological Chemistry, 278(32), 30403-30412. https://doi.org/10.1074/jbc.M304616200